The design and evaluation of a handheld digital microfluidic (DMF) bioagent analyzer is proposed by Core Microsolutions (CMSS) and University of Michigan Professor K.-D. Lee (College of Pharmacy) for a two year Phase I research period in the NIH Small Business Biodefense Program. This proposed research will integrate: 1) Electrowetting on Dielectric (EWOD) droplet-based microfluidic handling technology pioneered by CMSS Chairman C.-J. Kim and 2) unique bacteria, toxin capture/sensing methods developed by Professor Lee, for a complete point-of-care diagnostic device that is built upon a programmable, low cost, and generic microfluidic chassis that can be mass-manufactured for wide distribution in clinics and hospitals. A digital microfluidics (DMF) device is capable of highly programmable and elegant droplet (approximately 250 nl volume) generation, transport and splitting on a simple electrode array and is accomplished without the complex and costly micropump, microvalve, microchannel structures typically needed for lab-on-chip sensing devices. DMF handling also lowers reagent, sample consumption and opens up the possibility of signal amplification when a steady stream of droplets is directed to a single sensing site to increase target capture. Bacteria and toxin capture and sensing techniques developed by Professor Lee can be integrated directly on the EWOD electrode array and are hypothesized to provide bacteria and toxin detection with simple resistance or capacitance measurements. This Phase I feasibility of a DMF biosensor cartridge will be determined by evaluating limits of detection (LOD), signal/noise ratio, and dynamic range of several sample solutions with varying concentrations of bacteria and toxin targets. Precision and failure points of EWOD droplet handling will also be characterized. To accelerate the transition to Phase II commercial research, deliverables will include: a DMF biosensor cartridge, an optimized (2nd gen.) DMF microfluidics cartridge and an integrated EWOD controls module.